Method and composition for synchronizing time of insemination

ABSTRACT

Methods and compositions for synchronizing the time of insemination in swine are described. More particularly, methods are described for synchronizing the time of insemination by administration of a composition comprising a hormone, wherein the swine is inseminated only one time after administration of the hormone, and wherein there is no heat detection.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/153,973, filed May 13, 2016, which is a continuation of U.S.application Ser. No. 14/532,419, filed Nov. 4, 2014 (now issued U.S.Pat. No. 9,352,011), which is a continuation of U.S. application Ser.No. 13/265,307, filed Oct. 19, 2011 (now issued U.S. Pat. No.8,905,913), which is a U.S. national counterpart application, under 35U.S.C. § 3.71(c), of International Application Serial No.PCT/US2010/032258 filed Apr. 23, 2010, which claims priority under 35U.S.C. § 119(e) to U.S. Provisional Application No. 61/172,009, filedApr. 23, 2009, which are expressly incorporated by reference herein.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has been filedelectronically in ASCII format and is hereby incorporated by referencein its entirety. Said ASCII copy, created on Jul. 22, 2019, is named47191-267898_SL.txt and is 684 bytes in size.

FIELD OF THE INVENTION

The invention relates to methods and compositions for synchronizing thetime of insemination in an animal. More particularly, the inventionrelates to methods and compositions for synchronizing the time ofinsemination in a swine without heat detection.

BACKGROUND OF THE INVENTION

Gonadotropin-releasing hormone is a peptide of 10 amino acids and isalso known as luteinizing-hormone releasing hormone (LHRH).Gonadotropin-releasing hormone is produced in the hypothalamus, and isresponsible for the release of follicle-stimulating hormone andluteinizing hormone from the pituitary gland. Gonadotropin-releasinghormone is released from neurons in the hypothalmus, and plays a role inthe complex regulation of follicle-stimulating hormone and luteinizinghormone release. Follicle-stimulating hormone and luteinizing hormone,in combination, regulate the functioning of the gonads to producetestosterone in the testes and progesterone and estrogen in the ovaries,and regulate the production and maturation of gametes. For example,follicle-stimulating hormone stimulates the growth and recruitment ofimmature ovarian follicles in the ovary, and luteinizing hormonetriggers ovulation.

There are differences in gonadotropin-releasing hormone secretionbetween females and males. In males, gonadotropin-releasing hormone issecreted in pulses at a constant frequency, but in females the frequencyof the pulses varies during the estrus cycle and there is a large surgeof gonadotropin-releasing hormone just before ovulation.Gonadotropin-releasing hormone secretion is pulsatile in allvertebrates, and is necessary for correct reproductive function. Thus,gonadotropin-releasing hormone controls a complex process of folliculargrowth, ovulation, and corpus luteum maintenance in the female, andspermatogenesis in the male.

Gonadotropin-releasing hormone has been isolated and characterized as adecapeptide. Synthetic forms of gonadotropin-releasing hormone areavailable and modifications of the decapeptide structure ofgonadotropin-releasing hormone have led to multiplegonadotropin-releasing hormone analogs that either stimulate (e.g.,gonadotropin-releasing hormone agonists) or suppress (e.g.,gonadotropin-releasing hormone antagonists) the release of thegonadotropins, such as luteinizing hormone and follicle-stimulatinghormone.

It is important to commercial swine production to maximize reproductiveefficiency to make swine production more profitable. There has beenheavy reliance on daily heat detection of individual female swine withthe associated labor costs devoted to manual detection of heat in thefemale swine based on daily checks of gilts or sows to achieve the bestresults with, for example, artificial insemination. Heat detection usinglabor intensive methods, such as daily checks, increases the probabilityof success with artificial insemination. Thus, devoting time, manuallabor, and materials costs to daily checks for heat detection isnecessary because it is difficult to predict the time of heat (i.e., thebest time for insemination) without using methods requiring a dailyregimen for monitoring heat detection. Accordingly, methods are neededto optimize the success of insemination of animals, without heatdetection, to reduce the labor costs, and to increase the profitabilityof swine production.

SUMMARY OF THE INVENTION

Applicants have made the surprising discovery that controlling the timeof ovulation via hormone administration can eliminate breeding based onestrus detection and allow a swine to receive only one insemination foroptimal fertility and optimal cost expenditure. In one embodiment, amethod of synchronizing time of insemination in a swine without heatdetection is described. The method comprises the step of administeringto the swine, on the fourth day after weaning, a dose of a hormoneselected from the group consisting of a gonadotropin releasing hormone,a luteinizing hormone, a human chorionic gonadotropin, and combinationsthereof, wherein the swine is inseminated only one time about 15 toabout 24 hours after administration of the hormone and wherein there isno heat detection.

In the above described embodiment, the following features, or anycombination thereof, apply. In the above described embodiment, 1) theswine can be a postpartum sow; 2) the swine can be a gilt; 3) theinsemination can be an artificial insemination; 4) the insemination canbe through natural breeding; 5) ovulation can be synchronized; 6) thehormone can be administered in an amount effective to stimulate ovarianfollicle ovulation; 7) the dose of the hormone can be administered usinga method selected from the group consisting of use of a depositioncatheter, manual administration, and injection; 8) the swine can beinseminated about 15 to about 18 hours after administration of thehormone; 9) the swine can be inseminated about 18 hours afteradministration of the hormone; 10) the pregnancy rate of the swine canbe increased relative to a swine to which no hormone is administered;11) the litter size of the swine can be increased relative to a swine towhich no hormone is administered; 12) the hormone can be administered tomore than one swine; 13) the percentage of swine ovulating by about 48hours after administration of the hormone can be increased relative toswine to which no hormone is administered; 15) the hormone can beadministered intravaginally; 16) the hormone can be administered intothe anterior vagina; 17) the hormone can be a gonadotropin releasinghormone receptor agonist; 18) the hormone can be a luteinizing hormonereceptor agonist; 19) the hormone can be a human chorionic gonadotropinreceptor agonist; 20) the hormone can be triptorelin; 21) the hormonecan be a triptorelin salt; 22) the hormone can be synthetic; 23) thehormone can be in acetate form; 24) the hormone can be administered in acomposition comprising a gel; 25) the gel can comprise a polysaccharide;26) the polysaccharide can be selected from the group consisting ofcelluloses, dextrans, and alginates; 27) the gel can comprise acellulose; and 28) the cellulose can be methylcellulose. Any combinationof the embodiment of the preceding paragraph with 1-28, or anycombination thereof, is contemplated.

In another embodiment, a kit comprising a dose of a hormone selectedfrom the group consisting of a gonadotropin releasing hormone, aluteinizing hormone, a human chorionic gonadotropin, and combinationsthereof is described. The kit further comprises instructions for usewherein the hormone is in a composition comprising a gel and thecomposition has a pH of about 5 to about 6.

In the embodiment described in the preceding paragraph, the followingfeatures, or any combination thereof, apply. In the embodiment describedin the preceding paragraph, 1) the instructions can indicate that thehormone should be administered to a swine on about the fourth day afterweaning and that the swine can be inseminated one time about 15 to about24 hours after administration of the hormone; 2) the instructions canindicate that the insemination should be an artificial insemination; 3)the instructions can indicate that the insemination should be throughnatural breeding; 4) the kit can further comprise a deposition catheter,an applicator for manual administration, or a syringe; 5) theinstructions can indicate that the swine should be inseminated about 15to about 18 hours after administration of the hormone; 6) theinstructions can indicate that the swine should be inseminated about 18hours after administration of the hormone; 7) the instructions canindicate that the hormone should be administered intravaginally; 8) theinstructions can indicate that the hormone should be administered intothe anterior vagina; 9) the hormone can be a gonadotropin releasinghormone receptor agonist; 10) the hormone can be a luteinizing hormonereceptor agonist; 11) the hormone can be a human chorionic gonadotropinreceptor agonist; the hormone can be triptorelin; 12) the hormone can bea triptorelin salt; 13) the hormone can be synthetic; 14) the hormonecan be in acetate form; 15) the gel can comprise a saccharide; 16) thegel can comprise a polysaccharide; 17) the polysaccharide can beselected from the group consisting of celluloses, dextrans andalginates; 18) the gel can comprise a cellulose; and 19) the cellulosecan be methylcellulose. Any combination of the embodiment of thepreceding paragraph with 1-19, or any combination thereof, iscontemplated.

In another embodiment, a composition comprising a gonadotropin releasinghormone receptor agonist and a gel is described, wherein the compositionhas a pH of about 5.0 to about 6.0.

In the embodiment described in the preceding paragraph, the followingfeatures, or any combination thereof, apply. In the embodiment describedin the preceding paragraph, 1) the composition can further comprise apreservative; 2) the preservative can be selected from the groupconsisting of methylparaben and propylparaben; 3) the composition canfurther comprise a stabilizer; 4) the stabilizer can be an L-amino acid;5) the stabilizer can be L-methionine; 6) the gel can comprise apolysaccharide; 7) the polysaccharide can be selected from the groupconsisting of celluloses, dextrans and alginates; 8) the gel cancomprise a cellulose; 9) the cellulose can be methylcellulose; 10) thecomposition can further comprise a tonicity agent; 11) the agonist canbe triptorelin; 12) the composition can be combined with instructionsfor use; 13) the instructions in combination with the composition canindicate that a swine should be administered the agonist and that theswine should be inseminated one time about 15 to about 24 hours afteradministration of the agonist; and 14) the instructions can indicatethat the swine should be inseminated one time about 18 hours afteradministration of the agonist. Any combination of the embodiment of thepreceding paragraph with 1-14, or any combination thereof, iscontemplated.

The following various embodiments are provided.

1. A method of synchronizing time of insemination in a swine withoutheat detection is described. The method comprises the step ofadministering to the swine, on the fourth day after weaning, a dose of ahormone selected from the group consisting of a gonadotropin releasinghormone, a luteinizing hormone, a human chorionic gonadotropin, andcombinations thereof, wherein the swine is inseminated only one timeabout 15 to about 24 hours after administration of the hormone andwherein there is no heat detection.

2. The method of clause 1 wherein the swine is a postpartum sow.

3. The method of clause 1 wherein the swine is a gilt.

4. The method of clause 1 to 3 wherein the insemination is an artificialinsemination.

5. The method of clause 1 to 4 wherein the insemination is throughnatural breeding.

6. The method of clause 1 to 5 wherein ovulation is synchronized.

7. The method of clause 1 to 6 wherein the hormone is administered in anamount effective to stimulate ovarian follicle ovulation.

8. The method of clause 1 to 7 wherein the effective amount of thehormone is about 10 μg to about 1000 μg.

9. The method of clause 1 to 7 wherein the effective amount of thehormone is about 10 μg to about 500 μg.

10. The method of clause 1 to 9 wherein the dose of the hormone isadministered using a method selected from the group consisting of use ofa deposition catheter, manual administration, and injection.

11. The method of clause 1 to 10 wherein the swine is inseminated about15 to about 18 hours after administration of the hormone.

12. The method of clause 1 to 10 wherein the swine is inseminated about16 to about 24 hours after administration of the hormone.

13. The method of clause 1 to 10 wherein the swine is inseminated about18 to about 22 hours after administration of the hormone.

14. The method of clause 1 to 13 wherein the swine is inseminated about18 hours after administration of the hormone.

15. The method of clause 1 to 14 wherein the pregnancy rate of the swineis increased relative to a swine to which no hormone is administered.

16. The method of clause 1 to 15 wherein the total number of healthyfetuses is increased relative to a swine to which no hormone isadministered.

17. The method of clause 1 to 16 wherein the percentage farrowed isincreased relative to a swine to which no hormone is administered.

18. The method of clause 1 to 17 wherein the total number of pigletsborn is increased relative to a swine to which no hormone isadministered.

19. The method of clause 1 to 18 wherein the total number of pigletsborn per semen dose is increased relative to a swine to which no hormoneis administered.

20. The method of clause 1 to 19 wherein the total number of pigletsborn per semen dose is increased relative to a swine inseminatedfollowing heat detection.

21. The method of clause 1 to 20 wherein the piglet index is increasedrelative to a swine to which no hormone is administered.

22. The method of clause 1 to 21 wherein the piglet index is increasedrelative to a swine inseminated following heat detection.

23. The method of clause 1 to 22 wherein the litter size of the swine isincreased relative to a swine to which no hormone is administered.

24. The method of clause 1 to 23 wherein the hormone is administered tomore than one swine.

25. The method of clause 1 to 24 wherein the percentage of swineovulating by about 48 hours after administration of the hormone isincreased relative to swine to which no hormone is administered.

26. The method of clause 1 to 25 wherein the hormone is administeredintravaginally.

27. The method of clause 1 to 26 wherein the hormone is administeredinto the anterior vagina.

28. The method of clause 1 to 27 wherein the hormone is a gonadotropinreleasing hormone receptor agonist.

29. The method of clause 1 to 28 wherein the hormone is a luteinizinghormone receptor agonist.

30. The method of clause 1 to 29 wherein the hormone is a humanchorionic gonadotropin receptor agonist.

31. The method of clause 1 to 30 wherein the hormone is triptorelin.

32. The method of clause 1 to 31 wherein the hormone is a triptorelinsalt.

33. The method of clause 1 to 32 wherein the gonadotropin releasinghormone has the formula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH₂ (SEQID NO: 1).

34. The method of clause 1 to 33 wherein the gonadotropin releasinghormone receptor agonist has the formula

or a solvate, a hydrate, or a pharmaceutically acceptable salt thereof,wherein

R¹ and R² are independently in each instance hydrogen, or areindependently selected from the group consisting of alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl,and heteroarylalkyl, each of which is optionally substituted, or R¹ andR² and the attached carbon form a carbocycle or heterocycle;

X is hydrogen, or X is selected from the group consisting of, alkyl,cycloalkyl, heteroalkyl, optionally substituted alkylene-carboxamide;and

HNC(O)NR³R⁴, where R³ and R⁴ are in each instance independently selectedfrom the group consisting of hydrogen, alkyl, heteroalkyl and haloalkyl.

35. The method of clause 34 wherein R1 is methylene-heteroaryl, andwhere heteroaryl is selected from the group consisting of pyridyl,thiazolyl, pyridazolyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl,pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and

R2 is hydrogen or methyl.

36. The method of clause 34 to 35 wherein X is CH₂C(O)NH₂¬.

37. The method of clause 1 to 36 wherein the hormone is synthetic.

38. The method of clause 1 to 37 wherein the hormone is in acetate form.

39. The method of clause 1 to 38 wherein the hormone is administered ina composition comprising a gel.

40. The method of clause 39 wherein the gel comprises a polysaccharide.

41. The method of clause 40 wherein the polysaccharide is selected fromthe group consisting of celluloses, dextrans, and alginates.

42. The method of clause 39 to 41 wherein the gel comprises a cellulose.

43. The method of clause 42 wherein the cellulose is methylcellulose.

44. The method of clause 39 to 43 wherein the gel comprises about 0.25weight % to about 10 weight % of the methylcellulose.

45. The method of clause 39 to 43 wherein the gel comprises about 0.5weight % to about 4.0 weight % of the methylcellulose.

46. The method of clause 1 to 45 wherein the hormone is administeredwith a stabilizer, and wherein the stabilizer is L-methionine.

47. A kit comprising a dose of a hormone selected from the groupconsisting of a gonadotropin releasing hormone, a luteinizing hormone, ahuman chorionic gonadotropin, and combinations thereof, and instructionsfor use wherein the hormone is in a composition comprising a gel and thecomposition has a pH of about 5 to about 6 is described.

48. The kit of clause 47 wherein the instructions indicate that thehormone should be administered to a swine on about the fourth day afterweaning and that the swine should be inseminated one time about 15 toabout 24 hours after administration of the hormone.

49. The kit of clause 47 to 48 wherein the instructions indicate thatthe insemination should be an artificial insemination.

50. The kit of clause 47 to 49 wherein the instructions indicate thatthe insemination should be through natural breeding.

51. The kit of clause 47 to 50 wherein the kit further comprises adeposition catheter, an applicator for manual administration, or asyringe.

52. The kit of clause 47 to 51 wherein the instructions indicate theswine should be inseminated about 15 to about 18 hours afteradministration of the hormone.

53. The kit of clause 47 to 51 wherein the instructions indicate theswine should be inseminated about 16 to about 24 hours afteradministration of the hormone.

54. The kit of clause 47 to 51 wherein the instructions indicate theswine should be inseminated about 18 to about 22 hours afteradministration of the hormone.

55. The kit of clause 47 to 54 wherein the instructions indicate thatthe swine should be inseminated about 18 hours after administration ofthe hormone.

56. The kit of clause 47 to 55 wherein the instructions indicate thatthe hormone should be administered intravaginally.

57. The kit of clause 47 to 56 wherein the instructions indicate thatthe hormone should be administered in an effective amount of about 10 μgto about 1000 μg.

58. The kit of clause 47 to 56 wherein the instructions indicate thatthe hormone should be administered in an effective amount of about 10 μgto about 500 μg.

59. The kit of clause 47 to 58 wherein the instructions indicate thatthe hormone should be administered into the anterior vagina.

60. The kit of clause 47 to 59 wherein the hormone is a gonadotropinreleasing hormone receptor agonist.

61. The kit of clause 47 to 60 wherein the hormone is a luteinizinghormone receptor agonist.

62. The kit of clause 47 to 61 wherein the hormone is a human chorionicgonadotropin receptor agonist.

63. The kit of clause 47 to 62 wherein the hormone is triptorelin.

64. The kit of clause 47 to 63 wherein the hormone is a triptorelinsalt.

65. The kit of clause 47 to 64 wherein the gonadotropin releasinghormone has the formula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH₂ (SEQID NO: 1).

66. The kit of clause 47 to 65 wherein the gonadotropin releasinghormone receptor agonist has the formula

or a solvate, a hydrate, or a pharmaceutically acceptable salt thereof,wherein

-   -   R¹ and R² are independently in each instance hydrogen, or are        independently selected from the group consisting of alkyl,        heteroalkyl, cycloalkyl, heterocycloalkyl, haloalkyl, aryl,        heteroaryl, arylalkyl, and heteroarylalkyl, each of which is        optionally substituted, or R¹ and R² and the attached carbon        form a carbocycle or heterocycle;    -   X is hydrogen, or X is selected from the group consisting of,        alkyl, cycloalkyl, heteroalkyl, optionally substituted        alkylene-carboxamide; and    -   HNC(O)NR³R⁴, where R³ and R⁴ are in each instance independently        selected from the group consisting of hydrogen, alkyl,        heteroalkyl and haloalkyl.

67. The kit of clause 66 wherein R¹ is methylene-heteroaryl, and whereheteroaryl is selected from the group consisting of pyridyl, thiazolyl,pyridazolyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl, pyrrolyl,indolyl, benzopyrazolyl, and benzimidazolyl; and

R2 is hydrogen or methyl.

68. The kit of clause 66 to 67 wherein X is CH₂C(O)NH₂¬.

69. The kit of clause 47 to 68 wherein the hormone is synthetic.

70. The kit of clause 47 to 69 wherein the hormone is in acetate form.

71. The kit of clause 47 to 70 wherein the gel comprises a saccharide.

72. The kit of clause 47 to 71 wherein the gel comprises apolysaccharide.

73. The kit of clause 72 wherein the polysaccharide is selected from thegroup consisting of celluloses, dextrans and alginates.

74. The kit of clause 47 to 73 wherein the gel comprises a cellulose.

75. The kit of clause 74 wherein the cellulose is methylcellulose.

76. The kit of clause 47 to 75 wherein the gel comprises about 0.25weight % to about 10 weight % of the methylcellulose.

77. The kit of clause 47 to 75 wherein the gel comprises about 0.5weight % to about 4.0 weight % of the methylcellulose.

78. The kit of clause 47 to 77 further comprising a stabilizer whereinthe stabilizer is L-methionine.

79. A composition comprising a gonadotropin releasing hormone receptoragonist and a gel wherein the composition has a pH of about 5.0 to about6.0 is described.

80. The composition of clause 79 further comprising a preservative.

81. The composition of clause 79 to 80 wherein the preservative isselected from the group consisting of methylparaben and propylparaben.

82. The composition of clause 79 to 81 further comprising a stabilizer.

83. The composition of clause 82 wherein the stabilizer is an L-aminoacid.

84. The composition of clause 82 to 83 wherein the stabilizer isL-methionine.

85. The composition of clause 79 to 84 wherein the gel comprises apolysaccharide.

86. The composition of clause 85 wherein the polysaccharide is selectedfrom the group consisting of celluloses, dextrans and alginates.

87. The composition of clause 79 to 86 wherein the gel comprises acellulose.

88. The composition of clause 87 wherein the cellulose ismethylcellulose.

89. The composition of clause 79 to 88 wherein the gel comprises about0.25 weight % to about 10 weight % of the methylcellulose.

90. The composition of clause 79 to 88 wherein the gel comprises about0.5 weight % to about 4.0 weight % of the methylcellulose.

91. The composition of clause 79 to 90 further comprising a tonicityagent.

92. The composition of clause 79 to 91 wherein the agonist istriptorelin.

93. The composition of clause 79 to 92 in combination with instructionsfor use.

94. The composition of clause 93 wherein the instructions in combinationwith the composition indicate that a swine should be administered theagonist and that the swine should be inseminated one time about 15 toabout 24 hours after administration of the agonist.

95. The composition of clause 93 wherein the instructions in combinationwith the composition indicate that a swine should be administered theagonist and that the swine should be inseminated one time about 16 toabout 22 hours after administration of the agonist.

96. The composition of clause 93 wherein the instructions in combinationwith the composition indicate that a swine should be administered theagonist and that the swine should be inseminated one time about 18 toabout 22 hours after administration of the agonist.

97. The composition of clause 93 to 96 wherein the instructions indicatethat the swine should be inseminated one time about 18 to about 22 hoursafter administration of the agonist.

98. The composition of clause 79 to 97 wherein the gonadotropinreleasing hormone receptor agonist is gonadotropin releasing hormone,and wherein the gonadotropin releasing hormone has the formulapGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH₂ (SEQ ID NO: 1).

99. The composition of clause 79 to 98 wherein the gonadotropinreleasing hormone receptor agonist has the formula

or a solvate, a hydrate, or a pharmaceutically acceptable salt thereof,wherein

R¹ and R² are independently in each instance hydrogen, or areindependently selected from the group consisting of alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl,and heteroarylalkyl, each of which is optionally substituted, or R¹ andR² and the attached carbon form a carbocycle or heterocycle;

X is hydrogen, or X is selected from the group consisting of, alkyl,cycloalkyl, heteroalkyl, optionally substituted alkylene-carboxamide;and

HNC(O)NR³R⁴, where R³ and R⁴ are in each instance independently selectedfrom the group consisting of hydrogen, alkyl, heteroalkyl and haloalkyl.

100. The composition of clause 99 wherein R¹ is methylene-heteroaryl,and where heteroaryl is selected from the group consisting of pyridyl,thiazolyl, pyridazolyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl,pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and

R² is hydrogen or methyl.

101. The composition of clause 99 to 100 wherein X is CH₂C(O)NH₂¬.

102. The composition of clause 79 to 101 wherein the gonadotropinreleasing hormone receptor agonist is in an effective amount of about 10μg to about 1000 μg.

103. The composition of clause 79 to 101 wherein the gonadotropinreleasing hormone receptor agonist is in an effective amount of about 10μg to about 500 μg.

104. The composition of clause 79 to 103 comprising methylparaben,propylparaben, sodium chloride, sodium citrate, L-methionine, citricacid, triptorelin, and methycellulose.

105. The composition of clause 79 to 104 wherein the compositioncomprises methylparaben in an amount of about 0.09% weight per volume,propylparaben in an amount of about 0.01% weight per volume, sodiumchloride in an amount of about 0.91% weight per volume, sodium citratein an amount of about 0.186% weight per volume, L-methionine in anamount of about 0.1% weight per volume, citric acid in an amount ofabout 0.07% weight per volume, triptorelin in an amount of about 0.01%weight per volume, and methycellulose in an amount of about 1.2% weightper volume.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. shows the percentage of sows ovulating following treatments(0-hour) given at 96+/−2 hr post-weaning.

FIG. 2. shows triptorelin stability after 13 days at 0-4° C. (triangle),30° C. (square), and 50° C. (diamond).

DETAILED DESCRIPTION OF THE ILLUSTRATIVE EMBODIMENTS

In one embodiment, a method of synchronizing the time of insemination ina swine without heat detection is described. The method comprises thestep of administering to the swine, on the fourth day after weaning, adose of a hormone selected from the group consisting of a gonadotropinreleasing hormone, a luteinizing hormone, a human chorionicgonadotropin, and combinations thereof, wherein the swine is inseminatedonly one time about 15 to about 24 hours after administration of thehormone and wherein there is no heat detection.

In one embodiment, a kit comprising a dose of a hormone selected fromthe group consisting of, for example, a gonadotropin releasing hormone,a luteinizing hormone, a human chorionic gonadotropin, and combinationsthereof is described. In another embodiment, the kit further comprisesinstructions for use. In yet another embodiment, the hormone is in acomposition comprising a gel as herein described. The compositiontypically has a pH of about 5 to about 6, but the pH may range fromabout 4 to about 9.

In one embodiment, a composition comprising a gonadotropin releasinghormone receptor agonist is described. In another embodiment, thecomposition further comprising a gel is described. The compositiontypically has a pH of about 5 to about 6, but the pH may range fromabout 4 to about 9.

All of the illustrative embodiments, modifications, and alternativeforms described below may be applied to the embodiments described in thepreceding paragraphs of this Detailed Description section and to theembodiments described in the Summary of Invention.

The method for synchronizing the time of insemination in a swine withoutheat detection includes the step of administering to the swine, a doseof a hormone, for example, a gonadotropin releasing hormone, aluteinizing hormone, a human chorionic gonadotropin, derivatives oranalogs of gonadotropin-releasing hormone, luteinizing hormone, or humanchorionic gonadotropin, or combinations thereof. In accordance with oneembodiment, the hormone is administered to a swine. Any porcine species,e.g., gilts (i.e., female pigs prior to first mating), includingpubertal gilts, and sows, including postpartum sows, or any other typeof swine, may be used in the methods and may be administered thecompositions herein described.

Illustratively, swine are weaned on day 0 as herein described. Animalstypically receive a single dose of the hormone on day 4 post-weaning(i.e., the fourth day after weaning). Animals receiving treatment aretypically inseminated a single time at 15 hours (or 15 hours±2 hr), 16hours (or 16 hours±2 hr), 17 hours (or 17 hours±2 hr), 18 hours (or 18hours±2 hr), 19 hours (or 19 hours±2 hr), 20 hours (or 20 hours±2 hr),21 hours (or 21 hours±2 hr), 22 hours (or 22 hours±2 hr), 23 hours (or23 hours±2 hr), 24 hours (or 24 hours±2 hr), 27 hours (or 27 hours±2hr), or 30 hours (or 30 hours±2 hr) post hormone administration.

Breeding of the animal may be by any means of artificial insemination(AI), or through natural breeding. In any embodiment described herein, asecond breeding or subsequent breedings may be performed. In yet anotherembodiment, the swine is inseminated only one time. In anotherillustrative aspect, there is no heat detection. In another aspect,there is no heat detection between hormone administration and 48 hoursafter ovulation.

In any embodiment described herein, the hormone is administered, forexample, on the fourth day after weaning, i.e. about 96 hours afterweaning. In various illustrative embodiments, the hormone can beadministered, for example, about 80, about 82, about 84, about 86, about88, about 90, about 91, about 92, about 93, about 94, about 95, about96, about 97, about 98, about 99, about 100, about 102, about 104, orabout 108 hours after weaning. More typically, the hormone isadministered from about 94 to about 98 hours after weaning, i.e., about94, about 95, about 96, about 97 or about 98 hours after weaning. Inanother embodiment, the hormone is administered from about 92 to about106 hours after weaning.

In any embodiment described herein, the swine is inseminated one time,for example, about 15 to about 24 hours after administration of thehormone. In various further illustrative embodiments, the swine isinseminated about 15 to about 18 hours after administration of thehormone, about 13 to about 18 hours, about 15 to about 20 hours, orabout 13 to about 20 hours after administration of the hormone. In otherillustrative aspects, the swine is inseminated about 13 hours, about 14hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours,about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23hours, or about 24 hours after administration of the hormone.Illustratively, the swine is inseminated about 15 hours, about 18 hours,about 20 hours, about 22 hours, or about 24 hours after administrationof the hormone.

In any embodiment described herein, the pregnancy rate of the swine canbe increased relative to a swine to which no hormone is administered(e.g., to which a placebo is administered). For example, the pregnancyrate in hormone-treated animals can be about 75% to about 90%. Inanother embodiment, the litter size of the swine can be increasedrelative to a swine to which no hormone is administered (e.g., to whicha placebo is administered). For example, the typical litter size inhormone-treated animals can be about 20 to about 24 fetuses perpregnancy. In other embodiments, the litter size in hormone-treatedanimals can be about 14 to about 18, about 14 to about 24, or about 14to about 20 fetuses per pregnancy. In yet another embodiment, thepercentage of swine ovulating by about 48 hours after administration ofthe hormone can be increased relative to swine to which no hormone isadministered (e.g., to which a placebo is administered). For example,the percentage of animals that ovulate by about 48 hours afteradministration of the hormone can be about 65% to about 85%.

In further embodiments described herein, the total number of healthyfetuses can be increased relative to a swine to which no hormone isadministered. For example, the number of healthy fetuses forhormone-treated animals can be about 9 to about 18, about 9 to about 16,about 9 to about 14, or about 9 to about 12. In further embodimentsdescribed herein, the percent farrowed for hormone-treated animals canbe increased when compared to swine to which no hormone is administered.For example, the percentage farrowed for hormone-treated animals can beabout 76% to about 90%, about 76% to about 85%, or about 76% to about80%. In further embodiments described herein, the total number ofpiglets born to hormone-treated animals can be increased relative to aswine to which no hormone is administered. For example, the number ofpiglets born can be about 13 to about 18, about 13 to about 17, about 13to about 16, about 13 to about 15, or about 13 to about 14. In anyembodiment described herein, the pregnancy rate of the swine, the littersize of the swine, the total number of healthy fetuses, the farrowingpercent, and the total number of piglets born, for hormone-treatedanimals, can be similar to that of animals inseminated upon heatdetection.

In any embodiment described herein, the total number of piglets born persemen dose can be increased relative to a swine to which no hormone isadministered, and the total number of piglets born per semen dose can beincreased relative to animals inseminated upon heat detection. Forexample, the total number of piglets born ser semen dose can be about 6,about 7, about 8, about 9, about 10, about 11, about 12, about 14, about16, about 18, or about 20. In any embodiment described herein, thepiglet index (pigs born alive/100 sows allotted) can be increasedrelative to a swine to which no hormone is administered, and the pigletindex can be increased relative to animals inseminated upon heatdetection.

In any embodiment described herein, compositions for synchronizing thetime of insemination in a swine without heat detection comprise: a) ahormone; and b) a pharmaceutically acceptable pH buffering agent toprovide a pH in the range of about pH 4 to about pH 9. The pH of thecomposition herein described can range from about 4 to about 9. In otherembodiments, the pH can range from about 4 to about 8, from about 4 toabout 7, from about 4.5 to about 6.5, about 4.5 to about 6, or fromabout 5 to about 6.

Further, the hormone compositions can be produced, in accordance withthe dosage form, through a routine method by appropriately mixing with,diluting with, or dissolving in an additive such as various excipients,disintegrants, binders, salts, lubricants, local anesthetics (e.g.,lidocaine), diluents, preservatives, chelating agents, buffers, tonicityagents, antiseptic agents, wetting agents, emulsifiers, dispersants,stabilizers, a solution adjuvant, or combinations thereof.

Illustratively, the compositions comprising the hormone can be in theform of a gel and the composition can have, for example, a viscosity ofabout 10 (centipoise) cP to about 300,000 cP. In various illustrativeembodiments, the viscosity of the composition can be about 100 cP toabout 100,000 cP, about 250 cP to about 400 cP, about 300 cP to about400 cP, about 500 cP to about 100,000 cP, about 700 cP to about 100,000cP, about 200 cP to about 20,000 cP, about 200 cP to about 10,000 cP,about 200 cP to about 5,000 cP, about 200 to about 1,000 cP, about 200cP to about 600 cP, about 100 cP to about 600 cP, about 100 cP to about500 cP, about 200 cP to about 500 cP, about 200 cP to about 450 cP, orabout 100,000 cP to about 250,000 cP. In accordance with variousembodiments herein described, the viscosity of the composition can beabout 200 cP, about 250 cP, about 300 cP, about 400 cP, about 500 cP,about 1,000 cP, about 15,000 cP, about 20,000 cP, about 30,000 cP, about40,000 cP, about 50,000 cP, about 75,000 cP, about 100,000 cP, about200,000 cP, or about 300,000 cP. The viscosity of a solution can bemeasured using a viscometer, such as a rheometer, based on techniqueswell-known in the art.

Typically, the gels as described herein comprise about 0.001 to about3.0% weight/weight (w/w) of a hormone or a salt thereof, more typicallyabout 0.5-5.0% (w/w) or about 0.1-5.0% (w/w) of a hormone or a saltthereof, a preservative, a gel (i.e., a viscosity-modifying agent), abuffer to maintain a pH between about 5 to about 6, and a tonicity agentto maintain a tonicity between about 200 to about 400 mOsm/kG.

In accordance with any embodiment described herein, the composition issufficiently viscous that the composition stays adhered to the targettissue for a sufficient time to deliver an effective amount of thehormone. The typical viscosity will depend on factors such as, forexample, the rate of penetration of the hormone and the quantity of thehormone that is applied. Suitable viscosity modulating agents includebut are not limited to, ionic and non-ionic water soluble polymers;crosslinked acrylic acid polymers; hydrophilic polymers such aspolyethylene oxides, polyoxyethylene-polyoxypropylene copolymers, andpolyvinylalcohol; cellulosic polymers and cellulosic polymer derivativessuch as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropylmethylcellulose, hydroxypropyl methylcellulose phthalate, methylcellulose, carboxymethyl cellulose, and etherified cellulose; gums suchas tragacanth and xanthan gum; sodium alginate; gelatin, hyaluronic acidand salts thereof, chitosans, gellans or any combination thereof.

The viscosity modulating agent may be in the form of a gel, paste,cream, ointment, and the like. In one embodiment, the compositioncomprises a hormone and a gel, as a viscosity modifying agent, and thehormone is administered in the composition comprising the gel. In oneembodiment, the gel is a hydrogel, a lipogel, or a viscous sol. Inanother embodiment, the gel is a hydrogel. The gel may be prepared usingany method known in the art, for example, such as those methodsdescribed in U.S. Pat. Nos. 6,908,623 and 7,456,207, incorporated hereinby reference.

In any embodiment described herein, the gel (i.e., a viscosity modifyingagent) comprises a polysaccharide. In accordance with the methods andcompositions herein described, the polysaccharide may include, forexample, alginates and glucose, such as glycogens, starches (e.g.,amylose and amylopectin), celluloses, and dextrans. The polysaccharidecan be, for example, a methyl, ethyl, or propyl cellulose ester, ether,hydroxy-ether, hydroxy-alkyl, or hydroxy-ester. To achieve the desiredviscosity, a sufficient amount of one or more polysaccharides may beused. Typically, about 0.25 to about 10 weight % polysaccharide (basedon the total weight of the composition) is desirable. In anotherembodiment, the weight % of the polysaccharide is about 0.25 weight % toabout 3.0 weight %, about 1.0 weight % to about 7 weight %, about 1.0weight % to about 4.0 weight %, or about 1.0 weight % to about 2.0weight %. In other embodiments, the weight % of the polysaccharide isabout 0.1%, about 0.5%, about 0.75%, about 0.8%, about 0.9%, about 1.0%,about 1.1%, about 1.2%, about 1.4%, about 1.8%, about 2.0%, about 5%,about 8%, or about 10% (all in weight/weight). To increase the viscosityof the composition, the polysaccharide may be used in conjunction withone or more non-polysaccharide viscosifiers known in the art. Examplesof possible non-polysaccharide viscosifiers that could be used inconjunction with one or more polysaccharides include xantham gum,alginic acids and salts thereof, magnesium aluminum silicate, dextrins,sucrose and derivatives thereof, and mixtures thereof. The amount ofnon-polysaccharide viscosifier, if present, can be about 0.1 weight % toabout 10 weight %, depending on the desired viscosity.

In any embodiment described herein, the gel comprises a cellulose.Illustrative embodiments of the cellulose, as herein described, includemethylcellulose, ethylcellulose, hydroxypropyl cellulose, carbomethylcellulose, hydroxypropyl methyl cellulose, and hydroxyethyl methylcellulose. The cellulose can be a cellulose derivative, preferably anon-ionic cellulose ester, ether, hydroxy-ether, or hydroxy-ester, or anon-ionic starch derivative. Typically, about 0.25 weight % to about 10weight % of the cellulose (based on the total weight of the composition)is desirable. In another embodiment, the weight % of the cellulose isabout 0.25 weight % to about 3.0 weight %, about 0.5 weight % to about3.0 weight %, about 0.5 weight % to about 4.0 weight %, about 1.0 weight% to about 7 weight %, about 1.0 weight % to about 4.0 weight %, orabout 1.0 weight % to about 2.0 weight %. In other embodiments, theweight % of the cellulose is about 0.1%, about 0.5%, about 0.75%, about0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.4%, about1.8%, about 2.0%, about 5%, about 8%, or about 10% (all inweight/weight). If a uniform gel is desired, dispersing agents such asalcohol, sorbitol, or glycerin can be added, or the gelling agent can bedispersed by tituration, mechanical mixing, or stirring, or combinationsthereof.

Acceptable stabilizers for use in the described methods and compositionsinclude, an L-amino acid and an L-methionine. In other embodiments,stabilizers that can be used include, but are not limited to,polysaccharides such as acacia, agar, alginic acid, guar gum andtragacanth, gelatin and synthetic and semi-synthetic polymers such ascarbomer resins, cellulose ethers, and carboxymethyl chitin. Thestabilizer is generally in an amount of about 0.05 to about 10%, about0.05 to about 5%, about 0.05 to about 2.0%, about 0.05 to about 1.0%,about 0.05 to about 0.5%, about 0.05 to about 0.2%, about 0.1 to about5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1 to about 5%,about 1 to about 10%, about 1 to about 20% (all in weight/volume). Inone embodiment, in the presence of a stabilizer as herein described, theshelf life of the composition can be at least 12 months, at least 18months, or at least 24 months. In another embodiment, the compositioncan be stored at temperatures ranging from about 2° C. to about 8° C.Inert carriers can also be included such as lactose, starch, dextrin,dicalcium phosphate, and calcium sulfate. In one embodiment including astabilizer, the composition is chemically stable and remains at least99% pure, at least 99.5% pure, or at least 99.7% pure, for at leastthree months.

The tonicity agent can be non-ionic or ionic. Illustratively, acceptabletonicity agents for use in the described methods and compositionsinclude, for example, ionic agents such as sodium chloride, potassiumchloride, or a balanced salt solution. In accordance with oneembodiment, the tonicity agent is present in an amount to achieve atonicity between about 200-400 mOsm/kG, about 220-380 mOsm/kG, or about250-340 mOsm/kG. Non-ionic tonicity agents include diols, such asglycerol, mannitol, erythritol, polyethylene glycol, propylene glycol;and sugars such as sucrose and dextrose. The tonicity agent is generallyin an amount of about 0.05 to about 10%, about 0.05 to about 5%, about0.05 to about 2.0%, about 0.05 to about 1.0%, about 0.05 to about 0.5%,about 0.05 to about 0.2%, about 0.1 to about 5%, about 0.1 to about 10%,about 0.1 to about 20%, 0.5 to about 2.0%, about 0.6 to about 2.0%,about 0.5 to about 1.8%, about 0.6 to about 1.8%, about 1.0 to about5.0%, about 1.0 to about 10%, or about 1.0 to about 20% (all inweight/volume).

In any embodiment described herein, the pH buffering agents for use inthe compositions and methods herein described are those agents known tothe skilled artisan to be pH buffering agents or compositions andinclude, for example, acetate, borate, carbonate, citrate, and phosphatebuffers, as well as various biological buffers, for example, TAPS,Bicine, Tris, Tricine, HEPES, TES, MOPS, PIPES, Cacodylate, and MES.Other pH buffering agents include hydrochloric acid, sodium hydroxide,magnesium oxide, monopotassium phosphate, bicarbonate, ammonia, carbonicacid, sodium citrate, citric acid, acetic acid, disodium hydrogenphosphate, borax, boric acid, and the like. The buffering agent isgenerally in an amount of about 0.01 to about 10%, about 0.02 to about10%, about 0.02 to about 5%, about 0.02 to about 2.0%, about 0.02 toabout 1.0%, about 0.02 to about 0.5%, about 0.05 to about 10.0%, about0.05 to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%,about 0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%,about 1 to about 5%, about 1 to about 10%, about 1 to about 20% (all inweight/volume).

The buffering agent used in the formulations herein described can beused at any concentration needed to obtain the desired pH range. Forexample, the buffering agent can be used at a concentration of about0.001M to about 1M, about 0.001M to about 2M, about 0.001M to about 5M,about 0.05M to about 0.1M, about 0.05M to about 0.2M, about 0.05M toabout 1M, 0.05M to about 2M, about 0.05 to about 5M, about 0.1M to about1M, about 0.1M to about 2M, about 0.1M to about 5M. Any amount ofbuffering agent needed to obtain the desired pH range can be used in theformulations described herein. Typically, the pharmaceuticallyacceptable pH buffering agent can be used to provide a pH in the rangeof about pH 4 to about pH 9. The pH of the composition herein describedcan range from about 3 to about 10, or about 4 to about 9. In anyembodiment described herein, the pH can range from about 4 to about 8,from about 4 to about 7, from about 4.5 to about 6.5, about 4.5 to about6, from about 5 to about 6, about 5 to about 5.5, about 4 to about 6, orabout 4.5 to about 5.5.

In any embodiment described herein, the composition herein describedcomprises one or more pharmaceutically acceptable preservatives. As usedherein, the term “preservative” includes an agent or a combination ofagents that aids in stabilizing the composition, inhibiting microbialgrowth, or both. Examples of suitable preservatives include parabens(e.g., methyl, ethyl, propyl, and butyl esters of parahydroxybenzoicacid), propyl gallate, sorbic acid and its sodium and potassium salts,propionic acid and its calcium and sodium salts, “Dioxin”(6-acetoxy-2,4-dimethyl-m-dioxane), “Bronopol”(2-bromo-2-nitropropane-1,3-diol) and salicylanilides such asdisbromosalicylanilide, tribromosalicylamilides, “Cinaryl” 100 and 200or “Dowicil” 100 and 200 (Cis isomer of1-(3-chloroallyl-3,5,7-triaza-1-azanidadamantane chloride),hexachlorophene, sodium benzoate, citric acid, ethylenediaminetetraacetic acid and its alkali metal and alkaline earth metalsalts, butyl hydroxyanisol, butyl hydroxytoluene, phenolic compoundssuch as chloro- and bromocresols and chloro- and bromo-oxylenols,quaternary ammonium compounds like benzalkonium chloride, aromaticalcohols such as phenylethyl alcohol, benzyl alcohol, etc.,chlorobutanol, quinoline derivatives such as iodochlorohydroxyquinolin,and the like. The total amount of preservative, when present, is about0.005 weight % to about 2 weight %, about 0.001 weight % to 1.0 weight%, about 0.005 weight % to about 0.25 weight %, or about 0.05 weight %to about 0.2 weight %, typically about 0.01 weight % to about 0.1 weight% (all in weight/weight).

In any embodiment described herein, the pharmaceutical compositioncontains a chelating agent, such as those known to those skilled in theart, for example, ethylenediamine tetraacetate (EDTA),diethylenetriaminepentaacetic acid (DTPA), andN,N-bis(carboxymethyl)glycine (NTA), or salts thereof. The compositioncan contain about 0.003 weight % to about 1.0 weight %, about 0.02weight % to about 0.2 weight %, about 0.01 weight % to about 1.0 weight%, or about 0.02 weight % to about 0.5 weight % (all in weight/volume)of the chelating agent.

In any embodiment described herein, antimicrobial agents can be includedin the compositions described herein. Such agents may include, but arenot limited to 5-chloro-2-(2,4-dichlorophenoxy)-phenol,8-hydroxyquinoline, copper II compounds, phthalic acid, chlorhexidine,alexidine, hexetidine, sanguinarine, benzalkonium chloride,salicylanilide, domiphen bromide, cetylpyridinium chloride,tetradecylpyridinium chloride, N-tetradecyl-4-ethylpyridinium chloride,octenidine, iodine, sulfonamides, bisbiguanides, phenolics, delmopinol,octapinol, and other piperidino derivatives, and nicin preparations, anysuitable antibiotics such as augmentin, amoxicillin, tetracycline,doxycycline, minocycline, metronidazole, neomycin, kanamycin, andclindamycin, and any salts of any of these compounds where applicable,and any combinations of these compounds. In yet another embodiment,anti-fungal compounds can be included, alone or in combination with anyof the above-described antimicrobials. Anti-fungals agents that aresuitable for use in the compositions described herein include, but arenot limited to, nystatin, miconazole, econazole nitrate, clotrimazole,and flucytosine. The antimicrobial or anti-fungal agents can be added tothe formulations herein described in an amount of about 0.01 to about10%, about 0.01 to about 5%, about 0.01 to about 2.0%, about 0.01 toabout 1.0%, about 0.01 to about 0.5%, about 0.01 to about 0.2%, 0.05 toabout 10%, about 0.05 to about 5%, about 0.05 to about 2.0%, about 0.05to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1to about 5%, about 1 to about 10%, about 1 to about 20% (all inweight/volume).

In any embodiment described herein, antioxidants can also be added. Forexample, antioxidants used herein can include beta-carotene, vitamin E,vitamin C, vitamin A, tocopherol, butylated hydroxytoluene, butylatedhydroxyanisole, tertiary-butylhydroquinone, propyl gallate, ascorbicacid, sodium metabisulfite, uric acid, carotenoids, flavonoids,melatonin, and ethoxyquin. The antioxidants can be added to theformulations herein described in an amount of about 0.01 to about 10%,about 0.01 to about 5%, about 0.01 to about 2.0%, about 0.01 to about1.0%, about 0.01 to about 0.5%, about 0.01 to about 0.2%, 0.05 to about10%, about 0.05 to about 5%, about 0.05 to about 2.0%, about 0.05 toabout 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1to about 5%, about 1 to about 10%, about 1 to about 20% (all inweight/volume).

As described herein, the composition contains a hormone selected fromthe group consisting of, for example, gonadotropin releasing hormone,luteinizing hormone, human chorionic gonadotropin, derivatives andanalogs thereof, and combinations thereof, in an amount effective tosynchronize the time of insemination in a swine without heat detectionwhen used in the method described herein. Additional examples ofacceptable hormones for use in the methods and compositions describedherein include, prostaglandins, progestogens, progesterones, angrogens,testosterones, estrogens, estradiols, gonadotropins, derivatives andanalogs thereof, combinations thereof, and the like. The hormone can bein acetate form. Further, the hormone can be a gonadotropin-releasinghormone, luteinizing hormone, or human chorionic gonadotropin agonist ora gonadotropin-releasing hormone, luteinizing hormone, or humanchorionic gonadotropin antagonist. As used herein,“gonadotropin-releasing hormone” refers to any gonadotropin releasinghormone, including gonadotropin releasing hormone analogs andderivatives, and gonadotropin releasing hormone agonists andantagonists. In one embodiment, the gonadotropin releasing hormone canbe synthetic. In another embodiment, the gonadotropin-releasing hormonecan be GnRH (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH₂ (SEQ ID NO: 1))(see, for example, U.S. Pat. No. 5,688,506, incorporated herein byreference) or triptorelin. As used herein, “luteinizing hormone” refersto any luteinizing hormone, including luteinizing hormone analogs andderivatives, and luteinizing hormone agonists and antagonists. In oneembodiment, the luteinizing hormone can be synthetic. In anotherembodiment, the luteinizing hormone can be LH (see, for example, U.S.Pat. No. 5,444,167, incorporated herein by reference). As used herein,“human chorionic gonadotropin” refers to any human chorionicgonadotropin, including human chorionic gonadotropin analogs andderivatives, and human chorionic gonadotropin agonists and antagonists.In one embodiment, the human chorionic gonadotropin can be synthetic. Inanother embodiment, the human chorionic gonadotropin can be hCG (see,for example, U.S. Pat. Nos. 6,469,139, 4,400,316, and 4,804,626,incorporated herein by reference).

Examples of gonadotropin releasing hormone agonists for use hereininclude, but are not limited to, leuprolide, nafarelin, buserelin,[DAla⁶, des Gly-NH₂ ¹⁰]GnRH, [DLys⁶]GnRH, [DAla⁶] GnRH, [2-Me-Ala⁶]GnRH,[D-α-aminobutyroyl⁶, des-GlyNH₂ ¹⁰]GnRH, triptorelin, lutrelin,goserelin, deslorelin, and histrelin. Generally, gonadotropin releasinghormone agonists are modeled after the natural gonadotropin releasinghormone decapeptide with specific amino acid substitutions typically atpositions 6 and 10. Triptorelin is an example of a gonadotropinreleasing hormone agonist with only a single substitution at position 6.

Examples of gonadotropin releasing hormone antagonists include Antide (adecapeptide represented by the formulaD-Ac-D-2-Nal¹-DpClPhe²-D-3-Pal³-Ser4-NiLys⁵-D-NicLys⁶-Leu⁷-ILys⁸-Pro⁹-D-Ala¹⁰),[Ac-D4ClDPhe¹, D4ClDPhe², DTrp³, DArg⁶, DAla¹⁰]GnRH, [Ac-4ClDPhe²,D₃Pal³, Arg⁵, D₂Nal⁶, DAla¹⁰]GnRH, [Ac-D2-Na¹1, 4ClDPhe², DTrp³, DArg⁶,DAla¹⁰]GnRH, [Ac-D2 Nal¹, 4FDPhe², DTrp³, DArg⁶]GnRH, [Ac-D2Nal¹,4ClDPhe2, DTrp³, DhArg(Et₂)⁶, DAla¹⁰]GnRH, and [Ac—Na¹1, DME4ClPhe²,DPal³, Ser⁴, Tyr⁵, DArg⁶, Le⁷, ILys⁸, Pro⁹, DAla¹⁰]GnRH.

In any embodiment described herein, the use of a gonadotropin releasinghormone agonist of formula

or a solvate, a hydrate, or a pharmaceutically acceptable salt thereofis described wherein

R¹ and R² are independently in each instance hydrogen, or areindependently selected from the group consisting of alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl,and heteroarylalkyl, each of which is optionally substituted, or R¹ andR² and the attached carbon form a carbocycle or heterocycle; and

X is hydrogen, or X is selected from the group consisting of, alkyl,cycloalkyl, heteroalkyl, optionally substituted alkylene-carboxamide,and HNC(O)NR³R⁴, where R³ and R⁴ are in each instance independentlyselected from the group consisting of hydrogen, alkyl, heteroalkyl andhaloalkyl.

In another embodiment, the above use wherein R¹ is methylene-heteroaryl,where heteroaryl is selected from the group consisting of pyridyl,thiazolyl, pyridazolyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl,pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and R² ishydrogen or methyl is described.

In yet another embodiment, any one of the previously described useswherein X is CH₂C(O)NH₂ is described.

The gonadotropin releasing hormone agonsits and antagonists, and analogsthereof, such as the analogs described in the formula above, used hereincan be administered in the form of pharmaceutically acceptable non-toxicsalts or complexes. The salts include acid addition salts such as, forexample, hydrochloride, hydrobromide, sulfate, phosphate, nitrate,oxalate, fumarate, gluconate, tannate, maleate, acetate, benzoate,succinate, alginate, malate, ascorbate, tartrate and the like. Thecomplexes can be with metals such as for example zinc, barium, calcium,magnesium, aluminum and the like.

The amount of the hormone effective for use in accordance with themethods and compositions described herein depends on many parameters,including the molecular weight of the hormone, its route ofadministration, and whether it is in its native form. As in describedherein an “effective amount” of the hormone is an amount sufficient tosynchronize the time of insemination in a swine without heat detectionby using the method described herein. The effective amount of thehormone to be administered to a swine can range from about 10 μg toabout 2000 μg, about 10 μg to about 1000 μg, about 10 μg to about 500μg, about 10 μg to about 100 μg, about 10 μg to about 50 μg, about 50 μgto about 2000 μg, about 50 μg to about 1000 μg, about 50 μg to about 500μg, about 50 μg to about 300 μg, about 50 μg to about 200 μg, about 100μg to about 200 μg, about 100 μg to about 300 μg, about 100 μg to about500 μg, about 100 μg to about 1000 μg, about 200 μg to about 2000 μg, orabout 0.05 mg to about 50 mg. In various illustrative aspects, thehormone can be administered to a swine at a dose of about 20 μg, about50 μg, about 75 μg, about 100 μg, about 150 μg, about 180 μg, about 200μg, about 225 μg, about 250 μg, about 300 μg, about 400 μg, about 500μg, about 750 μg, about 1000 μg, about 1500 μg, or about 2000 μg of thehormone. The hormone can be administered in one or more doses.

The hormone in the composition described herein can be administered at aconcentration of, for example, about 50 μg/mL to about 500 μg/mL, about50 μg/mL to about 400 μg/mL, about 50 μg/mL to about 300 μg/mL, about 50μg/mL to about 200 μg/mL, about 50 μg/mL to about 150 μg/mL, about 50μg/mL to about 250 μg/mL, or about 100 μg/mL. The composition can beadministered in various volumes including for example a dosage volume of1 mL, 2 mL, 3 mL, 4 mL, or 5 mL.

In any embodiment described herein, the hormone is administered in anamount effective to stimulate ovarian follicle ovulation and tosynchronize ovulation according to the method described herein. The doseof the hormone can be administered using a method selected from thegroup consisting of 1) use of a deposition catheter, 2) manualadministration, 3) injection, or any other art recognized means foradministering a pharmaceutical composition, for example, any other artrecognized means for vaginally administering a pharmaceuticalcomposition, such as a composition containing a hormone. In oneembodiment, the hormone can be administered to more than one swine.

Examples of methods for effective hormone administration, other thanvaginal administration, include parenteral administration to the animal,for example, subcutaneously, intramuscularly, intraperitoneally,intrathecally, or intravenously, or in combination with an acceptablecarrier. Suitable means for parenteral administration include needle(including microneedle) injectors, needle-free injectors, and infusiontechniques. The parenteral compositions for use in accordance with thisinvention can be in the form of a reconstitutable lyophilizatecomprising one or more doses of the hormone composition. Examples ofparenteral dosage forms include aqueous solutions of the composition inwell-known acceptable liquid carriers such as liquid alcohols, glycols(e.g., polyethylene glycols), glucose solutions (e.g., 5%), esters,amides, sterile water, buffered saline (including buffers like phosphateor acetate; e.g., isotonic saline).

In any embodiment described herein, the composition can be administeredto the animal locally. Examples of local administration methods for useherein include, topical, intravaginal, and intrarectal. Examples ofdosage forms for use in this embodiment include creams, ointments, gels,pastes, powders, lotions, transdermal patches, intrauterine devices,vaginal rings, and vaginal tablets. In one illustrative embodiment, thecomposition is administered into the anterior vagina of the animal. Thecompounds may also be formulated in vaginal or rectal compositions suchas suppositories, e.g., containing conventional suppository bases suchas cocoa butter, carbowaxes, polyethylene glycols or other glycerides,all of which melt at body temperature, yet are solidified at roomtemperature.

The hormone may be administered to the animal by any useful proceduresand any effective dose and suitable dosage form can be used, includingoral dosage forms known in the art, such as pills, pellets, or capsules,and effective doses can be administered in standard or modified releasedosage forms. Modified release dosage formulations include delayed,sustained, pulsed, controlled, targeted, and programmed releaseformulations.

The compositions also may comprise suitable solid or gel phase carriersor excipients. Examples of such carriers or excipients include but arenot limited to calcium carbonate, calcium phosphate, various sugars,starches, cellulose derivatives, gelatin, and polymers such aspolyethylene glycols.

In another illustrative aspect of the invention, a kit is provided. Thekit comprises a dose or multiple doses of a hormone as described herein.In this embodiment, the kit can further comprise an applicator formanual administration, a deposition catheter, and/or a syringe forapplication of the hormone composition to an animal. In yet anotherembodiment, the hormone is in a composition comprising a gel asdescribed herein. In one illustrative embodiment, the kit may comprisethe hormone and the gel separately for mixing before administration tothe animal. In another embodiment, the kit may comprise the hormone andthe gel admixed in a vessel for immediate administration.

In yet another embodiment, the kit contains instructions for use. Theinstructions may indicate that the insemination should be throughnatural breeding; the insemination should be through artificialinsemination; the swine should be inseminated at about 15 to about 18,about 18 to about 22, or about 15 to about 24 hours after administrationof the hormone; or the swine should be inseminated at about 15, about16, about 17, about 18, about 19, about 20, about 21, about 22, about23, about 24, or about 30 hours after administration of the hormone.Other suitable kit components include excipients, disintegrants,binders, salts, lubricants, local anesthetics (e.g., lidocaine),diluents, preservatives, chelating agents, buffers, tonicity agents,antiseptic agents, wetting agents, emulsifiers, dispersants,stabilizers, and the like. These components may be available separatelyor admixed with the hormone and/or gel as necessary. Any of the hormoneembodiments and any of the composition embodiments described herein canbe used to formulate the kit.

In yet another embodiment, an article of manufacture is provided. Thearticle of manufacture can comprise any of the compositions describedherein. The composition can be in a primary container, for example, aglass vial, such as an amber glass vial with a rubber stopper and/or analuminum tear-off seal. In another embodiment, the primary container canbe plastic or aluminum, and the primary container can be sealed. Inanother embodiment, the primary container may be contained within asecondary container to further protect the composition from light. Thesecondary container can be, for example, cardboard. Any of theseembodiments also apply to the kit embodiments described above, and anyof the hormone and composition embodiments described herein can apply tothe article of manufacture.

EXAMPLES Example 1 Study Design and Treatment Groups

Approximately 120 parity one to seven sows (30 sows within eachtreatment group) were used to obtain ovulation and reproductive tractdata. All sows were of the same genotype (PIC C22). Following 16 to 24days of lactation, sows were blocked by length of lactation and parityand randomly assigned to one of four treatments: 1) a placebo group,receiving a placebo gel at 96±2 hours post-weaning and a single AI 24hours later, 2) triptorelin gel at 96±2 hours post-weaning and AI 18 to20 hours later. 3) triptorelin gel at 96±2 hours post-weaning and AI 24to 26 hours later, and 4) triptorelin gel at 96±2 hours post-weaning andAI 30 to 32 hours later. The placebo formulation was identical to theactive formulation but without the hormone being present. All treatmentswere deposited into the anterior vagina within 2 cm of the cervix.Follicular development and ovulation was monitored via real timeultrasound at 24, 32, 40, 48, and 56 hours after treatment. Thirty (30)days after insemination, all sows were slaughtered and reproductivetracts obtained. Pregnancy rates and embryo survival were determined.

Example 2 Test Substance

The active ingredient was triptorelin(pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH₂ (SEQ ID NO: 1)) suppliedin the acetate form (molecular weight: 1371.6), from Bachem, Torrance,Calif. (Item H-4075 CGMP grade). Triptorelin gel (200 μg/2 mL), lot054023833-1, was formulated at Chem Laboratories Ltd (Auckland, NewZealand) in a gel composed of 1.2% Methocel™ Premium A4000 (DowChemicals) in sodium citrate, pH 5.5 with methyl and propyl paraben,NaCL, EDTA and L-methionine. Formulation vehicle, lot 054006645-1, wasformulated at Chem Laboratories Ltd. and was composed of 1.2% Methocel™Premium A4000 (Dow Chemicals) in sodium citrate, pH 5.5 with, NaCl andmethyl and propyl paraben. Fifty milliliters of triptorelin gel (200 μgtriptorelin acetate/mL) or formulation vehicle were packaged in a 50 mLAmber Borosilicate Glass Serum Vials (610206-50) with a Gray ButylPharmaceutical Serum Vial Stopper (73828A-SS) with a Standard AluminumSeal (SAS20NAT). The test substances were stored refrigerated (about 5°C.) and transported in insulated containers with appropriate ice packs.

Example 3 Example Formulations

Example formulations for the composition described in this applicationare shown in Tables 1 and 2.

TABLE 1 Weight Ingredient Function (% w/v) Methylparaben, sodium salt(USNF) Anti-microbial preservative 0.0900 Propylparaben, sodium salt(USNF) Anti-microbial preservative 0.0100 Sodium chloride, laboratoryreagent Tonicity agent 0.910 Sodium citrate, dihydrate Buffering agent0.186 L-Methionine, laboratory reagent Stabilizing agent 0.100 Citricacid, anhydrous Buffer 0.0700 Triptorelin acetate Active Pharmaceutical0.0100 Ingredient (API) Water (USNF) Dissolving solvent 98.4Methylcellulose (A4M Premium) Thickening agent 1.20 (USP)

TABLE 2 Amount per Quality 100 mg Component Standard Function % w/wTriptorelin Acetate In house Drug Substance 11.0 mg 0.011%* PurifiedWater USP Solvent 97.6 g 97.54%* Methylparaben, NF Preservative 89.0 mgSodium Salt** 0.089%* Propylparaben, NF Preservative 10.0 mg Sodium Salt** 0.010*    Sodium Chloride USP Tonicity agent 901 mg 0.901% L-Methionine USP Stabilizing agent 99.0 mg 0.099%  Sodium Citrate USPBuffering agent 184 mg 0.184%  Citric Acid USP Buffering agent 69.0 mg0.069%* Methycellulose USP Viscosity modifier 1.1 g  1.10%* *Nominalamount **Tested to compendial standard

Example 4 Estrus Observation

Sows were housed in gestation crates following weaning (Day 0). Boarswere housed in separate rooms, and/or at least 12 m away and downwind.To determine onset and duration of estrus, sows were observed for estrusdaily (1) from Day 3 until the end of estrus was confirmed, or (2) untilDay 6, which ever came first. To elicit signs of estrus, a mature boarwas walked slowly in the alley in front of the crates of the sows,exposing each test sow to visual, auditory and olfactory signals fromthe boar for up to 5 min. In keeping with standard practice atcommercial farms, while the boar was near the front of the sow's crate,estrus was tested by an experienced person applying back pressure to themidsection of the sow combined with side rubbing. Estrus was confirmedwhen a sow stood rigidly to the back pressure, with no vocalization andwith some indication of an ear reflex.

Example 5 Administration and Insemination

Triptorelin was administered at 96 hours (±2 hr) post-weaning as asingle 2 mL dose deposited approximately 1-2 cm posterior to the cervixwith a modified artificial insemination catheter. Weaning of sowsoccurred between 10 a.m. and noon on Day (O). The timing of theinsemination for each group of sows was given as directed in theprotocol. Those sows scheduled to be inseminated 18 hours followingadministration of triptorelin were generally inseminated between 6:00a.m. and 8:00 a.m. the following day, those sows scheduled to beinseminated 24 hours following administration of triptorelin weregenerally inseminated between noon and 2:00 p.m. the following day andthose sows scheduled to be inseminated 30 hours following administrationof triptorelin were generally inseminated between 6:00 p.m. and 8:00p.m. the following day. The single insemination for each sow wasperformed as they were normally done at the farm.

Example 6 Ovulation and Estrus Detection

All sows were observed for ovulation by transrectal ultrasonography.Ultrasonography was performed at 24 hr (±1.5 hour) after treatment onDay 5 and again at 8:00 p.m. (±1.5 hour) on Day 5. On Day 6 transrectalultrasonography was performed at 4:00 a.m. (±1 hour), noon (±1 hr) and8:00 p.m. (±1 hr) until ovulation was complete or until the 8:00 p.m.ultrasonography on Day 6 (the 56 hour mark from the animals treated at96 hours post-weaning) whichever came first. An Aloka 500 ultrasoundmachine was used for this purpose, with a 7.5 MHz linear arraytransducer attached to a fixed-angle PVC stabilizing rod to facilitateinsertion into the rectum. The transducer and PVC rod were coated with agynecological lubricant and gently inserted into the rectum until theovaries could be visualized, one at a time. The diameters of the threelargest follicles were to be recorded (to the nearest 0.1 mm) at eachscanning. A sow was declared to have ovulated when the number of largefollicles 6.5 mm) fell to less than 3.

On Day 5 post-weaning, the average size of the largest follicle was 7.3mm and was not influenced by treatment. Estrus expression within 7 daysof weaning averaged 79% and did not differ significantly amongtreatments. There was also no effect of treatment on the interval fromweaning to estrus (average 112 hours).

The percentage of sows that had ovulated was not significantly affectedby treatment, at 24 hours, 32 hours or 40 hours following treatment (seeTable 3 and FIG. 1). However, the percentage of sows ovulating by 48hours after treatment was increased (p=0.0054) in all triptorelintreated groups [74.0% (AI at 18 hours), 77.0% (AI at 24 hours), and76.8% (AI at 30 hours)] compared to vehicle treated sows (42.6%).Treatment did not affect the percentage of sows ovulating by 56 hoursafter treatment (p=0.10), however sows treated with triptorelin had ahigher percentage of sows ovulating (average of 80.4%) than sows treatedwith vehicle (58.3%). While the average wean to ovulation interval didnot differ between triptorelin treated and vehicle treated animals, agreater synchrony of ovulation was observed in triptorelin treated sowswith 70-77% of sows ovulating during their most synchronous 24 hr period(120 to 144 hr after weaning) compared to vehicle treated sows with only39.9% of sows ovulating during their most synchronous period (128 to 156hours after weaning).

Example 7 Pregnancy Rate and Litter Size

Pregnancy rates, calculated as the percentage of animals allotted toeach treatment group that were pregnant at slaughter, averaged 66.6% anddid not differ significantly among treatment groups although pregnancywas numerically highest in triptorelin treated sows inseminated 18 hoursfollowing treatment (74.4%) followed by triptorelin treated sowsinseminated at 24 hours following treatment (68.1%), triptorelin treatedsows inseminated at 30 hours following treatment (62.3%) and placebotreated sows inseminated at 24 hours following treatment (61.8%). Therewas a tendency (p=0.09) for triptorelin treated sows inseminated at 18hours (12.4 fetuses) and at 24 hours (11.8 fetuses) post treatment tohave a higher number of healthy fetuses than triptorelin treated sowsinseminated at 30 hours (8.6) post treatment and for vehicle treatedsows inseminated at 24 hours (8.6) following treatment. The weight ofhealthy fetuses did not differ significantly among the treatment groups.

TABLE 3 Least squares means for response variable. Placebo OG OG OG AI24 AI 18 AI 24 AI 30 SEM P N 31 32 32 32 Estrus (%) 81.5 81.5 78.3 75.77.50 0.94 Wean to Estrus 114.6 114.3 108.6 111.8 4.04 0.69 Interval (h)Average Day 5 7.1 7.3 7.2 7.3 0.18 0.85 Follicle Size (mm) Wean to 140.0140.3 139.9 139.3 1.63 0.97 Ovulation Interval (h) Interval from AI19.6^(x) 26.3^(y) 19.9^(x) 13.3^(z) 1.63 0.0001 to Ovulation (h) OV24(%) 2.7 2.7 0 5.8 3.14 0.58 OV32 (%) 15.3 8.9 12.1 13.4 5.95 0.89 OV40(%) 31.3 27.9 31.0 27.7 8.19 0.98 OV48 (%) 42.6^(x) 74.0^(y) 77.0^(y)76.8^(y) 8.00 0.0054 OV56 (%) 58.3 77.2 82.2 81.7 7.74 0.10 Pregnant (%)61.8 74.4 68.1 62.3 8.07 0.66 Number of 8.6 12.4 11.8 8.6 1.40 0.09Healthy Fetuses Weight of 1.5 1.6 1.6 1.6 0.04 0.17 Healthy Fetuses (g)Means with differing superscripts within a row differ significantly at p= 0.05 Abbreviations are as follows: N (number of sows), Estrus (estrusduring the period of Day 3 until Day 6), OV24 (percent sows ovulated at24 hr after treatment), and Pregnant (pregnant sows of all sowsallotted. AIOVh Std err: 1.8379 1.5804 1.5483 1.5700 # Healthy FetusesStd err: 1.3966 1.3966 1.3968 1.4105

Example 8 Study 1—United Feeds Research Farm

Three study replicates were performed at an 800 sow research farm. Allreplicates included estrus and ovulation data and two replicatesincluded AI, pregnancy and litter results. At weaning, mixed parity sows(n=32/treatment) received either vehicle carrier at 96 h post-weaningand a single AI 24 h later (Placebo), the test substance OVUGEL™ at 96 hpost-weaning and a single AI 18 h later (OG18), OVUGEL™ at 96 hpost-weaning and a single AI 24 h later (OG24), or OVUGEL™ at 96 hpost-weaning and a single AI 30 h later (OG30). OVUGEL™ was administered˜2 cm posterior to the cervix using the deposition catheter. Estrusdetection was performed once daily from d 3 after weaning until d 7.Ultrasound was performed every 8 h from d 5 through d 6. On d 30 ofgestation, the sows were sacrificed and reproductive tracts examined.

There was no effect of treatment on sows expressing estrus within 7 daysof weaning (79%) or on the wean-to-estrus interval (4.7 days). Thepercentage of sows that ovulated by 48 h after treatment was greater(P=0.005) in OG18, OG24, and OG30 than in the Placebo treatment (Table4). The interval from AI to ovulation was different among treatmentgroups (P<0.001, Table 4) and occurred furthest from ovulation for OG18,intermediate for Placebo and OG24, and closest for OG30. Pregnancy rateswere not influenced by treatment (P=0.60, Table 4) but were influencedby interval from AI to ovulation (P=0.02). There was a significanteffect of treatment (P=0.003) on the number of healthy fetuses at day 30with more fetuses in OG18 and OG24 compared to the OG30 and Placebotreatments (Table 4). There was no effect of treatment on number of CL(P=0.30) but there was a tendency (P=0.09) for an effect on embryosurvival (Table 4).

Results indicate that OVUGEL™ effectively synchronized ovulation withoutaffecting pregnancy rate, and that litter size at day 30 of pregnancywas greater following a single AI at 18 or 24 hours after OVUGEL™compared to AI at 30 h after OVUGEL™ or compared to single inseminationsafter Placebo treatment.

TABLE 4 Unadjusted means for treatment responses. Placebo OG18 OG24 OG30SE Ovulate by 48 h, % 40.6^(a) 71.9^(b) 75.0^(b) 75.0^(b) 4.2 AI* toOvulation (h) 20.3^(a) 26.4^(b) 19.1^(a) 15.5^(c) 1.0 Pregnant (%) 68.286.4 81.8 68.2 4.6 Fetuses 12.9^(a) 16.5^(b) 17.4^(b) 12.6^(a) 0.6 CLnumber 18.5 22.1 23.0 17.6 1.2 ES (%) 53.2 65.9 67.3 57.0 2.4 AI =Artificial Insemination CL = Corpora Lutea ES = Embryo Survival OG =OVUGEL ™

Example 9 Study 2—Bache Farm Study

Sows were weaned (Day 0) and screened for enrollment in the study, bodycondition recorded, and assigned to a treatment group. Postpartum sows,from parity 1 to 10, were blocked by lactation length, parity, and bodycondition score. No sows with a body condition score below 2.5 of above3.5 was used. Genotypes used were those typically used in commerce inthe U.S. and Canada. Treatment groups were stalled in areas separatefrom each other so that those sows bred later in the day did not receivemultiple sessions of boar exposure. Each treatment sow received a singledose of the triptorlein composition (OVUGEL™) on Day 4 post-weaning.Control sows were untreated.

Estrus detection was performed on Day 4, and continued once a day until4 days after treatment (Day 8) or until sows no longer showed estrus.Control sows (untreated) were inseminated at intervals followingdetection of estrus as is normally practiced at the farm. OVUGEL™treated sows were inseminated with a single insemination at 15 hours(6:00 am+/−0.5 hr), 18 hours (9:00 am+/−0.5 hr), 21 hours (12:00pm+/−0.5 hr), 24 hours (3:00 pm+/−0.5 hr), and 27 hours (6:00 pm+/−0.5hr) post treatment.

Estrus detection was performed once daily between Days 18-24 followinginsemination to determine if the sows had recycled. Sows that havereturned to estrus were recorded and then rebred following normal farmSOP (these animals were off the study but subject to the withdrawalperiod). Trans-abdominal ultrasound (once) to confirm pregnancy in sowsthat had not returned to estrus was performed 28 to 32 days afterinsemination. The withdrawal period was verified 51 days post treatment.The date of farrowing, number of live born pigs and number of stillbornpigs within 24 hours after birth were recorded. Piglets were fosteredaccording to usual farm protocols.

At least 90 sows were allocated to each of the following groups.Controls were untreated. All other sows were given OVUGEL™ containing200 μg triptorelin, as the acetate.

-   -   1) Controls: Untreated and inseminated at intervals following        detection of estrus as is normally practiced on each site;    -   2) Triptorelin: Inseminated once at 15 hr (+/−0.5 hrs) after        triptorelin treatment;    -   3) Triptorelin: Inseminated once at 18 hr (+/−0.5 hrs) after        triptorelin treatment;    -   4) Triptorelin: Inseminated once at 21 hr (+/−0.5 hrs) after        triptorelin treatment;    -   5) Triptorelin: Inseminated once at 24 hr (+/−0.5 hrs) after        triptorelin treatment;    -   6) Triptorelin: Inseminated once at 27 hr (+/−0.5 hrs) after        triptorelin treatment.

TABLE 5 Time of Insemination Following OvuGel Administration Bred 15Bred 18 Bred 21 Bred 24 Bred 27 Bred hrs hrs hrs hrs hrs Contrast, FarmPost Post Post Post Post Std Err Treatment SOP OvuGel ™ OvuGel ™OvuGel ™ OvuGel ™ OvuGel ™ Means P< Returns 19.46^(ab) 794 ^(b)    799^(b)     19.72 ^(ab) 16.18 ^(ab) 22.74 ^(a) 4.5420 0.09 At 21 d, %Pregnant 79.28^(bc) 80.80 ^(c) 82.55 ^(c) 60.66 ^(a) 71.08 ^(abc) 64.52^(ab) 5.5913 0.03 At 28 d, % % Farrow 75.83 ^(b) 77.32 ^(b) 79.11 ^(b)59.44 ^(a) 67.63 ^(ab) 59.44 ^(a) 5.7977 0.04 Of Wean Total 12.2711.84   13.10   12.39   12.15 12.59 0.5563 0.65 Born Born 11.15 10.19  11.48   10.84   10.41 11.13 0.6030 0.62 Alive Stillborn 1.13  1.65   1.62    1.56   1.74 1.47 0.3780 0.88 Mummies 0.51 ^(b)   0.28 ^(ab)  0.27 ^(ab)  0.11 ^(a) 0.36 ^(ab) 0.40 ^(ab) 0.1110 0.21 Data werecollected from 366 sows (61 sows per treatment). LS means calculatedusing the PROC GLM procedure of SAS and assumed a model with treatmentand cohort. There was no treatment by cohort interaction for any of thevariables tested. LS means with different superscripts differ at P <0.05.

Example 10 Hormone Stability

FIG. 2 shows triptorelin stability (250 μg per mL⁻¹). The effect of pHwas analyzed at 0-4° C., 30° C., and 50° C. (Mcllvaine buffer). FIG. 2shows an optimal pH range of about 5 to about 6 for hormone stability.

Example 11 Stabilizer Analysis

The stability of formulations containing various stabilizers werestudied at a 3-month time point. The data summarized in Table 6indicates that L-Methionine provided the most stable formulation.

TABLE 6 Stability summary of formulations containing stabilizers TotalRel % % TA Subs Formulation Condition Label (as peak (each StabilizerF-0 (° C.) Claim area) >0.15%) None 11 25 94% 98% 2.6% (Control) 50 43%71% 28.8% EDTA 17 2-8 98% 98% 0.5% 0.02% 25 94% 97% 1.6% 50 78% 89%10.7% L-methionine 18 2-8 102%  99% None (0.1%) >0.3% 25 99% 98% 0.7% 5087% 90% 9.3% EDTA/L- 23 2-8 101%  98% 0.7% methionine 25 98% 98% 0.7%(0.02/0.1%) 40 91% 94% 4.7% 50 79% 90% 8.9% Mannitol 25 2-8 96% 98% 0.7%(3%) 40 89% 94% 5.6% 50 76% 87% 11.0% EDTA/L- 26 2-8 98% 99% 0.4%methionine/ 40 88% 94% 5.0% D-Mannitol 50 78% 88% 11.4% (0.02/0.1/3%)EDTA/L- 27 2-8 95% 96% 0.3% methionine/D- 40 86% 95% 4.4% Mannitol/P-8050 73% 88% 10.9% (0.02/0.1/3/0 .1%) Where P-80 as polysortrate 80

Example 12 Method of Preparing Hormone Composition

In one method of preparation embodiment, the hormone is dissolved in avessel containing citric acid and water, but separate from the otheringredients. The other ingredients are then added to the hormonecomposition, and water is added before addition of the methylcellulosecomposition. The methylcellulose composition is added slowly with highshear mixing to ensure homogeneity of the composition and to avoidclumping.

Example 13 Timed Insemination Following Intravaginal GNRH AgonistTreatment in PostPartum Sows

This study shows the effect of a single, fixed timed inseminationfollowing GnRH administration on subsequent farrowing rate and littersize as compared to control sows bred at onset of estrus and daily forthe duration of estrus (Table 7 and 8). One hundred ninety-nine weanedsows (PIC) were blocked by parity (parities 1 through 6; average parity2.9), previous lactation length (range 13 to 23 d; average length 19 d),and body condition score (range 2.0 to 4.0; average score 3.1) andallocated to one of two treatments. Control sows were observed forbehavioral estrus for 7 d after weaning and inseminated the day theywere observed in estrus and at 24 h intervals for the duration ofestrus. Sows in the GnRH treatment group (GnRH sows) were administeredintravaginally with the GnRH agonist preparation (OvuGel™), described inExamples 2 and 3, in the morning four days after weaning and inseminatedonce 21-22 h after OvuGel™ treatment. The GnRH sows were also observedfor 7 d after weaning for signs of behavioral estrus but wereinseminated without regard to signs of standing estrus.

Of the 99 Control sows, 90.9% (90) were bred by 7 d post-weaningcompared to 100% (100) of the GnRH sows (P<0.01). Control sows averaged2.1 inseminations per sow while GnRH sows all had 1 insemination per sow(P<0.01). There was no difference (P>0.75) in number of sows farrowingbetween Control and GnRH sows, 79.2% (79 sows) and 80.7% (81 sows),respectively. There was no effect of treatment on number of stillborns(P>0.45) and mummies (P>0.15). GnRH sows farrowed an average of 11.2pigs born alive compared to 11.4 pigs born alive to Control sows(P>0.65). Pigs born per semen dose was 5.2 vs. 9.6 (P<0.01) for Controland GnRH sows, respectively. These data indicate that treating sows withOvuGel™ and inseminating once following treatment results in farrowingrates and litter sizes comparable to sows receiving multipleinseminations during behavioral estrus, and a higher number of pigs bornper semen dose for the OvuGel™-treated animals.

TABLE 7 Responses to treatment and comparisons between Control and GnRHsows ¹ Control GnRH Number of Sows Allotted 99 100 Wean to EstrusInterval, d 4.4 4.3 In Estrus at Insemination, % 100 90 No. Inseminatedby 7 d post-weaning 90 100 Doses of Semen per Sow Inseminated 2.1 1.0Returns at 21 d of Sows Inseminated, % 6.7 8 No. Returns at 21 d 6 8Pregnant at 30 d of Sows Allotted, % 81.8 85 No. Pregnant at 30 d 81 85No. Sows Farrowed 79 81 Sows Farrowed of Sows Allotted, % 79.8 81.0Total Born per Litter 12.1 11.8 Total Born per Semen Dose 5.3 9.6 BornAlive per Litter 11.4 11.2 Stillborn per Litter 0.7 0.6 Mummies perLitter 0.8 0.6 Piglet Index (Pigs Born Alive/100 Sows Allotted) 906 909¹ GnRH sows were treated between 99 and 102 hours following weaning andinseminated between 21 and 22 hours post treatment. Control sows wereuntreated and bred normally. Numbers in the table are unadjusted meansand raw numbers.

TABLE 8 Responses to treatment (LS means ± SEM) and comparisons betweenControl and GnRH sows¹ Contrast, Control GnRH SEM P< Sows Allocated toTreatments 99 100 * * Sows Inseminated in 7-d period after 90 100 * *weaning Sows In Estrus at AI (%) 100 90.0 2.28 0.005 Weaning-to-.EstrusInterval (d) 4.4 4.3 0.06 0.33 No. of Inseminations/Sow 2.1 1.0 0.020.0001 Sows Pregnant of Sows Allotted (%) 81.4 84.8 3.74 0.52 No. SowsFarrowed 79 81 * * Sows Farrowed of Sows Allotted (%) 79.2 80.7 3.940.78 Total Pigs Born/Semen Dose 5.2 9.6 0.45 0.0001 Total Pigs BornAlive (all litters) 901 910 * * Total Pigs Born/Litter 12.1 11.8 0.360.56 Pigs Born Alive/Litter 11.4 11.2 0.34 0.67 ¹Data collected from 199sows (99 Control, 100 GnRH). Control sows were bred following normalfarm SOP. GnRH sows were treated on d 4 post-weaning and inseminatedonce 21-22 hrs later. LS means calculated using the PROC MIX procedureof SAS and assumed a model with treatment and cohort. There was notreatment by cohort interaction for any of the variables tested.

Example 14 Timed Insemination Following GNRH Agonist Admnistration inWeaned Sows

This study shows the effect of a single, fixed timed inseminationfollowing GnRH administration on subsequent farrowing rate and littersize (Tables 9 and 10). Three hundred weaned sows (PIC) were blocked byparity (parities 1 through 6; average parity 2.8), previous lactationlength (range 17 to 25 d; average length 21 d), and body condition score(range 2.5 to 3.5; average score 2.8) and allocated to one of twotreatments. Control sows were observed for 7 d after weaning forbehavioral estrus and inseminated the day they were observed in estrusand at 24 h intervals for the duration of estrus. Sows in the GnRHtreatment group (GnRH sows) were also observed for 7 d after weaning forsigns of behavioral estrus but were treated intravaginally with the GnRHagonist preparation (OvuGel™), described in Examples 2 and 3, on d 4after weaning and then inseminated once 24±2 h post-OvuGel™ treatment,regardless of whether or not they showed signs of standing estrus. Ofthe 150 Control sows, 80.7% (121) were bred by 7 d post-weaning comparedto 100% (150) of the GnRH sows (P<0.01).

Control sows averaged 2.3 inseminations per sow while GnRH sows all had1 insemination per sow (P<0.01). There was no difference (P>0.40) innumber of sows farrowing between Control and GnRH sows, 72.7% (109 sows)and 76.7% (115 sows), respectively. There was no effect of treatment onnumber of stillborns (P>0.30) and mummies (P>0.45). GnRH sows farrowedan average of 11.3 pigs born alive compared to 10.9 pigs born alive toControl sows (P<0.37). Pigs born per semen dose was 5.6 vs. 9.6 (P<0.01)for Control and GnRH sows, respectively. These data indicate thattreating sows with the GnRH agonist, OvuGel™ and inseminating once withrespect to the time of GnRH treatment results in farrowing rates andlitter sizes comparable to sows receiving multiple inseminations duringbehavioral estrus. The results also show that a higher amount of pigsare born per semen dose for the OvuGel™-treated animals than the controlanimals.

TABLE 9 Responses to treatment and comparisons between Control and GnRHsows ¹ Control GnRH Number of Sows Allotted 150 150 Wean to EstrusInterval, d 4.4 4.4 In Estrus at Insemination, % 100 84 No. Inseminatedby 7 d post-weaning 121 150 No. Inseminated 8 to 15 d post-weaning 5 .Doses of Semen per Sow Inseminated 2.2 1.0 Returns at 21 d of SowsInseminated, % 3.3 8.7 No. Returns at 21 d 4 13 Pregnant at 30 d of SowsAllotted, % 74.0 80.0 No. Pregnant at 30 d 111 120 No. Sows Farrowed 109115 Sows Farrowed of Sows Allotted, % 72.7 76.7 Total Born per Litter12.2 12.6 Total Born per Semen Dose 5.6 9.6 Born Alive per Litter 10.911.3 Stillborn per Litter 1.0 0.9 Mummies per Litter 0.3 0.4 PigletIndex (Pigs Born Live/100 Sows Allotted) 794 867 ¹ Sows were treated at96 hours following weaning and inseminated 24 ± 2 hours later on d 5post-weaning (120 to 122 hr post-weaning). Control sows were untreatedand bred normally. Numbers in the table are unadjusted means and rawnumbers.

TABLE 10 Responses to treatment (LS means ± SEM) and comparisons betweenControl and GnRH sows ¹ Contrast, Control GnRH SEM P< Sows Allocated toTreatments 150 150 * * Sows Inseminated in 7-d period after 121 150 * *weaning Sows In Estrus at AI (%) 100 84.0 2.34 0.0001 Weaning-to-EstrusInterval (d) 4.4 4.4 0.05 0.75 No. of Inseminations/Sow 2.3 1.0 0.040.0001 Sows Pregnant of Sows Allotted (%) 74.0 80.0 3.44 0.22 No. SowsFarrowed 109 115 * * Sows Farrowed of Sows Allotted (%) 72.7 76.7 3.560.43 Total Pigs Born/Semen Dose 5.6 9.6 0.43 0.0001 Total Pigs BornAlive (all litters) 1191 1300 * * Total Pigs Born/Litter 12.2 12.6 0.310.41 Pigs Born Alive/Litter 10.9 11.3 0.29 0.37 ¹ Data collected from300 sows (150 Control, 150 GnRH). Control sows were bred followingnormal farm SOP. GnRH sows were treated on d 4 post-weaning andinseminated once 24 +/− 2 hrs later. LS means calculated using the PROCMIX procedure of SAS and assumed a model with treatment and cohort.There was no treatment by cohort interaction for any of the variablestested.

Example 15 Statical Analysis

These data were analyzed using the Proc MIXED and General linear modelsprocedures of SAS (Cary, N.C.) for continuous and discrete variables.For all analyses, the models contained the fixed effects of treatment(placebo controls inseminated 24 hours after treatment, triptorelintreated sows inseminated 18 hours after treatment, triptorelin treatedsows inseminated 24 hours after treatment and triptorelin treated sowsinseminated 30 hours after treatment) and replicate. The records for sowparity and lactation length were included as covariates. First orderinteraction of treatment and replicate was tested and removed whennon-significant. Differences between treatments were tested on leastsquares means estimates using the T test at P<0.05.

What is claimed is:
 1. A method of synchronizing time of insemination ina swine, the method comprising the steps of, administering to the swine,on the fourth day after weaning, a dose of a hormone of the formula

or a solvate, a hydrate, or a pharmaceutically acceptable salt thereof,wherein R¹ and R² are independently in each instance hydrogen, or areindependently selected from the group consisting of alkyl, heteroalkyl,cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl,and heteroarylalkyl, each of which is optionally substituted, or R¹ andR² and the attached carbon form a carbocycle or heterocycle; X ishydrogen, or X is selected from the group consisting of, alkyl,cycloalkyl, heteroalkyl, optionally substituted alkylene-carboxamide;and HNC(O)NR³R⁴, where R³ and R⁴ are in each instance independentlyselected from the group consisting of hydrogen, alkyl, heteroalkyl andhaloalkyl, and inseminating the swine wherein the swine is inseminatedonly one time about 15 to about 24 hours after administration of thehormone, and wherein there is no heat detection at the time ofinsemination.
 2. The method of claim 1 wherein the swine is a postpartumsow.
 3. The method of claim 1 wherein the insemination is an artificialinsemination.
 4. The method of claim 1 wherein the hormone isadministered in an effective amount and the effective amount of thehormone is about 10 μg to about 500 μg.
 5. The method of claim 1 whereinthe dose of the hormone is administered using a method selected from thegroup consisting of use of a deposition catheter, manual administration,and injection.
 6. The method of claim 1 wherein the swine is inseminatedabout 18 to about 22 hours after administration of the hormone.
 7. Themethod of claim 1 wherein the pregnancy rate of the swine and/or thenumber of healthy fetuses is increased relative to a swine to which nohormone is administered.
 8. The method of claim 1 wherein the hormone isadministered into the anterior vagina.
 9. The method of claim 1 whereinthe hormone is a triptorelin salt.
 10. The method of claim 1 wherein thehormone is a gonadotropin releasing hormone and the hormone has theformula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH₂ (SEQ ID NO: 1). 11.The method of claim 1 wherein the hormone is in acetate form.
 12. Themethod of claim 1 wherein the hormone is administered in a compositioncomprising a gel.
 13. The method of claim 12 wherein the gel comprises apolysaccharide.
 14. The method of claim 13 wherein the polysaccharide isa cellulose and the cellulose is methylcellulose.
 15. The method ofclaim 14 wherein the gel comprises about 0.5 weight % to about 4.0weight % of the methylcellulose.
 16. The method of claim 1 wherein thehormone is administered with a stabilizer, and wherein the stabilizer isL-methionine.
 17. The method of claim 1 wherein the swine is inseminatedintravaginally.
 18. The method of claim 1 wherein the hormone issynthetic.
 19. The method of claim 1 wherein the hormone is administeredto more than one swine.